Characterization of extrachromosomal circular DNAs in plasma of patients with clear cell renal cell carcinoma.

BACKGROUND AND PURPOSE: Extrachromosomal circular DNAs (eccDNAs) have been recognized for their significant involvement in numerous biological processes. Nonetheless, the existence and molecular characteristics of eccDNA in the peripheral blood of patients diagnosed with clear cell renal cell carcinoma (ccRCC) have not yet been reported. Our aim was to identify potentially marked plasma eccDNAs in ccRCC patients. METHODS AND MATERIALS: The detection of plasma eccDNA in ccRCC patients and healthy controls was performed using the Tn5-tagmentation and next-generation sequencing (NGS) method. Comparisons were made between ccRCC patients and healthy controls regarding the distribution of length, gene annotation, pattern of junctional nucleotide motif, and expression pattern of plasma eccDNA. RESULTS: We found 8,568 and 8,150 plasma eccDNAs in ccRCC patients and healthy controls, respectively. There were no statistical differences in the length distribution, gene annotation, and motif signature of plasma eccDNAs between the two groups. A total of 701 differentially expressed plasma eccDNAs were identified, and 25 plasma eccDNAs with potential diagnostic value for ccRCC have been successfully screened. These up-regulated plasma eccDNAs also be indicated to originate from the genomic region of the tumor-associated genes. CONCLUSION: This work demonstrates the characterization of plasma eccDNAs in ccRCC and suggests that the up-regulated plasma eccDNAs could be considered as a promising non-invasive biomarker in ccRCC.

Ultrasound-Microwave Combined Extraction of Novel Polysaccharide Fractions from Lycium barbarum Leaves and Their In Vitro Hypoglycemic and Antioxidant Activities.

Ultrasound-microwave combined extraction (UMCE), gradient ethanol precipitation, chemical characterization, and antioxidant and hypoglycemic activities of Lycium barbarum leaf polysaccharides (LLP) were systematically studied. The optimal conditions for UMCE of LLP achieved by response surface method (RSM) were as follows: microwave time of 16 min, ultrasonic time of 20 min, particle size of 100 mesh, and ratio of liquid to solid of 55:1. Three novel polysaccharide fractions (LLP30, LLP50, LLP70) with different molecular weights were obtained by gradient ethanol precipitation. Polysaccharide samples exhibited scavenging capacities against ABTS and DPPH radicals and inhibitory activities against α-glucosidase and α-amylase. Among the three fractions, LLP30 possessed relatively high antioxidant and hypoglycemic activities in vitro, which showed a potential for becoming a nutraceutical or a phytopharmaceutical for prevention and treatment of hyperglycemia or diabetes.

Overexpression of Spondin-2 Is Associated with Recurrence-Free Survival in Patients with Localized Clear Cell Renal Cell Carcinoma.

BACKGROUND: The spondin-2 (SPON2) gene is overexpressed in multiple malignant tumors and may promote tumor aggressiveness. However, its expression profile and functional roles in clear cell renal cell carcinoma (ccRCC) are still unclear. METHODS: SPON2 expression in ccRCC was evaluated using expression data from TCGA and GEO databases, then confirmed by local patient population (94 patients). The clinical significance of SPON2 expression was evaluated. Downregulation of SPON2 was performed using small-interfering RNA (siRNA). The effects of SPON2 silencing on cell proliferation, apoptosis, invasion, and migration in vitro were investigated. RESULTS: SPON2 was overexpressed in the majority of the ccRCC at both mRNA and protein levels. SPON2 expression was significantly correlated with stage, grade, and recurrence (all P < 0.05) in patients with localized ccRCC. The receiver operating characteristic (ROC) curve showed that SPON2 expression could serve as a predictor of recurrence. SPON2 expression was significantly associated with recurrence-free survival (RFS) in patients with localized ccRCC. Knocking down SPON2 resulted in suppressed cell invasion and migration in vitro. CONCLUSION: SPON2 expression might function as a prognostic biomarker in patients with localized ccRCC.

A Randomized, 8-Week Study of the Effects of Extended-Release Paliperidone and Olanzapine on Heart Rate Variability in Patients With Schizophrenia.

PURPOSE: This study aimed to explore the effect of extended-release paliperidone (paliperidone ER) and olanzapine on heart rate variability (HRV) in patients with schizophrenia. METHODS: A total of 106 patients with schizophrenia diagnosed by the Diagnostic and Statistical Manual of Mental Disorders (Fifth Edition) were randomly divided into the paliperidone ER group or the olanzapine group for an 8-week clinical trial, with 53 patients in each group. The time domain and frequency domain analyses including the SD of all the R-R intervals in 24 hours (SDNN), the SD of the mean value of all the normal R-R intervals in every 5-minute interval within 24 hours (SDANN index), the mean value of the SD of all the normal R-R intervals in every 5-minute interval within 24 hours (SDNN index), the root mean square of successive R-R differences, the percentage of adjacent R-R intervals that differ by more than 50 milliseconds, high-frequency power (HF), low-frequency power (LF), and LF/HF were adopted to assess the HRV of patients at baseline and after treatment for 8 weeks in each group. The Positive and Negative Symptom Scale was used to evaluate the clinical efficacy. The incidence rates of adverse reactions were also calculated. RESULTS: In total, 48 patients in the paliperidone ER group and 45 patients in the olanzapine group completed the entire 8-week treatment. The SDNN, SDNN index, and SDANN index in the olanzapine group were significantly lower than those in the paliperidone ER group (P < 0.05) after treatment for 8 weeks, whereas their mean LF level was higher than that in the paliperidone ER group (P < 0.05) after completion of treatment. Patients in the olanzapine group showed a significant decrease in the SDNN, SDANN index, and SDNN index as well as a statistical increase in the LF and LF/HF in comparison with the pretreatment values (P < 0.05), whereas patients in the paliperidone ER group showed a decrease in the SDANN index and a statistical increase in the LF in comparison with the pretreatment values (P < 0.05). CONCLUSION: The HRV of patients with schizophrenia changes when they are administered with paliperidone ER or olanzapine, and more attention should be paid to their cardiac autonomic function when using these 2 antipsychotics.

Overexpression of COL5A1 promotes tumor progression and metastasis and correlates with poor survival of patients with clear cell renal cell carcinoma.

BACKGROUND AND AIMS: COL5A1 has been identified to be involved in metastasis of clear cell renal cell carcinoma (ccRCC) by bioinformatic analysis. This study aimed to investigate COL5A1 expression and its clinical significance in ccRCC. The function of COL5A1 in ccRCC was further investigated. METHODS: COL5A1 expression was examined in 256 ccRCC tissues and paired adjacent normal renal tissues by immunohistochemistry and real-time quantitative PCR. The clinical significance of COL5A1 expression was evaluated. Downregulation of COL5A1 was achieved using siRNA. The effects of COL5A1 silencing on cell proliferation, apoptosis, migration, invasion in vitro, and tumor growth in vivo were investigated. RESULTS: COL5A1 expression was upregulated in the majority of the ccRCC tissues at both protein and mRNA levels. COL5A1 expression was significantly correlated with tumor diameter, tumor stage, tumor grade, distant metastasis, recurrence, necrosis, and sarcomatoid (all P<0.05). COL5A1 expression was also significantly associated with overall survival of ccRCC patients (HR 1.876; P=0.027) and recurrence-free survival of localized ccRCC patients (HR 4.751; P<0.001). The accuracy of TNM, University of California Los Angeles Integrated Staging System, and Mayo clinic stage, size, grade, and necrosis prognostic models was improved when COL5A1 expression was added. CONCLUSION: COL5A1 knockdown significantly inhibited cell proliferation, induced cell apoptosis, inhibited cell migration and invasion in vitro, and inhibited tumor growth in vivo. Therefore, COL5A1 may be a novel prognostic biomarker and a promising therapeutic target for ccRCC.

Room-temperature borylation and one-pot two-step borylation/Suzuki-Miyaura cross-coupling reaction of aryl chlorides.

A highly efficient room-temperature borylation strategy of aryl chlorides is described. Utilizing Buchwald's second-generation preformed catalyst, boronate esters were obtained for a wide range of substrates in high yield. The method was also applied to Suzuki-Miyaura cross-coupling reaction in a one-pot two-step sequential manner, providing a facile and convenient access to the direct synthesis of biaryl compounds from aryl chlorides.

Cobalt oxide 2D nano-assemblies from infinite coordination polymer precursors mediated by a multidentate pyridyl ligand.

In this work, the construction of Co3O4 two dimensional (2D) nano-assemblies utilizing infinite coordination polymers (ICPs) as precursors was investigated, aiming at the morphology targeted fabrication and utilization of 2D materials. Based on the successful modulation of morphology, a rose-like Co based ICP precursor was obtained, which was further transformed into porous Co3O4 nanoflake assemblies with a well-preserved 2D morphology and a large surface area. The mechanism of the morphology modulation was illustrated by systematic investigation, which demonstrated the crucial role of a modulating agent in the formation of 2D nano-assemblies. In addition, the cobalt oxide 2D nano-assemblies are fabricated into a lithium anode combined with graphene, and the remarkable capacity and stability (900 mA h g(-1) after 50 cycles) of the resulting Co3O4/G nanocomposite indicates its potential in lithium battery applications.

H2O2 inhibits proliferation and mediates suppression of migration via DLC1/RhoA signaling in cancer cells.

BACKGROUND: RhoGTPase-activating proteins (RhoGAPs) regulate RhoGTPases in cells, but whether individual reactive oxygen species (ROS) regulate RhoGAPs is unknown. Our previous published papers have shown that deleted in liver cancer 1 (DLC1) inhibits cancer cell migration by its RhoGAP activity. The present study was designed to explore the role of H2O2 in regulation of DLC1. MATERIALS AND METHODS: We treated cells with H2O2 for 24h and phenotypic changes were analyzed by MTT, RT-PCR, Western blotting, immunofluorescence staining and wound healing assays. RESULTS: H2O2 downregulated cyclin D1 and cyclin E to inhibit proliferation, and upregulated BAX to induce apoptosis in MCF-7 cells. Compared with non-tumorigenic cells, H2O2 increased expression of DLC1 and reduced activity of RhoA in cancer cells. Stress fiber production and migration were also suppressed by H2O2 in MDA-MB-231 cells. CONCLUSIONS: Our study suggests that H2O2 inhibits proliferation through modulation of cell cycle and apoptosis-related genes, and inhibits migration by decreasing stress fibers via DLC1/RhoA signaling.

Zn(II)-benzotriazolate clusters based amide functionalized porous coordination polymers with high CO2 adsorption selectivity.

Two new porous coordination polymers (PCPs) based on different nanosized C3 symmetry ligands and Zn(II)-benzotriazolate clusters have been synthesized solvothermally. Both of the desolvated complexes show selective uptake of CO2 over CH4 and N2 at ambient temperature.

[Forensic psychiatric cases of 534 mental disorders caused by traumatic brain injury].

OBJECTIVE: To analyze the types of mental disorders caused by traumatic brain injury and the optimal time for forensic psychiatric appraisal. To explore the relationship between the degree of traumatic brain injury, the time of appraisal and the grade of intellectual deficiency. METHODS: Five hundred and thirty-four forensic psychiatric cases of mental disorders caused by traumatic brain injury were retrospectively analyzed. RESULTS: In the types of mental disorders caused by traumatic brain injury, the most cases were diagnosed as organic mood disorders (51.1%), following organic neurosis-like syndrome (24.0%) and organic intellectual deficiency (18.0%). For the disability grades, the most cases were the level VIII and IX disability grades, 219 cases (41.0%) and 177 cases (33.1%), respectively. The degree of brain injury and the degree of intelligence defection according to WAIS-RC were higher in intellectual deficiency group compared with non-intellectual deficiency group (P < 0.05). The grade of disability correlated with the degree of brain injury, the result of WAIS-RC, the result of cerebral CT scanning, and the grade of brain electrical activity mapping (BEAM) abnormality (P < 0.05). Nevertheless, the degree of intellectual deficiency did not correlate with appraisal time and the degree of brain injury (P > 0.05). CONCLUSION: The factors influencing intellectual deficiency are complex. The findings of objective examination including cerebral CT scanning, BEAM, WAIS-RC and others should be considered as important indexes for disability evaluation.

In vivo experimental testing of the FW axial blood pump for left ventricular support in Fu Wai Hospital.

A fully implantable, axial flow blood pump has been developed in Fu Wai Hospital aiming for clinical use. This ventricular assist device (VAD), which was developed after numerous CFD analyses for the flow characteristics of the pump, is 58.5-mm long, 30-mm wide (including DC motor), and weighs 240 g. The pump can deliver 5 L/min for pressures of 100 mm Hg over 8,000 rpm. In this study, short-term hemocompatibility effects of the axial left ventricular assist device (LVAD) (FW blood pump) were evaluated in four healthy sheep. The device was implanted into the left ventricular apex of beating hearts. The outflow graft of each device was anastomosed to the descending aorta. The hemolysis, which was evaluated in vivo by free hemoglobin value, was below 30 mg/dL. Evaluation of serum biochemical data showed that implantation of the FW blood pump in sheep with normal hearts did not impair end organ function. Gross and microscopic sections of kidney, liver, and lung revealed no evidence of microemboli. Performance of the pump in vivo was considered sufficient for a LVAD, although further design improvement is necessary in terms of hemolysis and antithrombosis to improve biocompatibility of the pump.

Homologous recombination as an evolutionary force in the avian influenza A virus.

Avian influenza A viruses (AIVs), including the H5N1, H9N2, and H7N7 subtypes, have been directly transmitted to humans, raising concerns over the possibility of a new influenza pandemic. To prevent a future avian influenza pandemic, it is very important to fully understand the molecular basis driving the change in AIV virulence and host tropism. Although virulent variants of other viruses have been generated by homologous recombination, the occurrence of homologous recombination within AIV segments is controversial and far from proven. This study reports three circulating H9N2 AIVs with similar mosaic PA genes descended from H9N2 and H5N1. Additionally, many homologous recombinants are also found deposited in GenBank. Recombination events can occur in PB2, PB1, PA, HA, and NP segments and between lineages of the same/different serotype. These results collectively demonstrate that intragenic recombination plays a role in driving the evolution of AIVs, potentially resulting in effects on AIV virulence and host tropism changes.

Homologous recombination is apparent in infectious bursal disease virus.

Infectious bursal disease virus (IBDV) is a non-enveloped double-stranded RNA virus belonging to the Birnaviridae family. It shows substantial variation in the major antigen region of the viral capsid protein VP2, where a hypervariable region plays a key role in the virulence of IBDV and its epitope. This study identifies several putative recombinants from previously published data to suggest that homologous recombination may naturally occur between different IBDV strains. In addition, a novel very virulence sublineage emerges in the VP2 phylogenic tree, comprising three putative recombination strains isolated in Korea and China, KSH, KK1 and SH-h. The major putative parents of the three mosaics are descended from the vaccine lineage while their hypervariable regions from vvIBDV. These findings also suggest that vaccine coverage may have influence on the evolution and genetic diversity of IBDV, resulting in a novel group with vvIBDV phenotype through recombination with wild IBDV.

Homologous recombination evidence in human and swine influenza A viruses.

Dynamic gene mutation and the reassortment of genes have been considered as the key factors responsible for influenza A virus virulence and host tropism change. This study reports several significant evidence demonstrating that homologous recombination also takes place between influenza A viruses in human and swine lineages. Moreover, in a mosaic descended from swine H1N1 subtype and human H2N2, we found that its minor putative parent might be a derivative from the human cold-adapted vaccine lineage, which suggests that live vaccine is capable of playing a role in genetic change of influenza A virus via recombination with circulating viruses. These results would be important for knowing the molecular mechanism of mammal influenza A virus heredity and evolution.

Design optimization of an axial blood pump with computational fluid dynamics.

A fully implantable, axial flow blood pump has been developed in our hospital. Both in vitro and in vivo tests showed that the hemolysis and thrombus characteristics of the pump were in an acceptable but not in an ideal range. Computational fluid dynamics (CFD) and in vitro test results showed that the pump worked at off-design point with a low hydraulic efficiency; CFD analysis also showed regions of reverse flow in the diffuser, which not only decreases the pump's hydrodynamic efficiency, but also increases its overall potential for blood trauma and thrombosis. To make a blood pump atraumatic and nonthrombogenic, several methods were taken to reach a final model of the optimized blood pump using CFD, which decreased the rotational speed from 9,000 to 8,000 rpm, and the design flow rate from 11 to 6 L/min. More significantly, the flow separation and recirculation in the diffuser region were eliminated, which mitigated the traumatic and thrombus effect on blood. The acceptable results of the numerical simulations encourage additional in vitro and in vivo studies.

Development of the functionally total artificial heart using an artery pump.

Based on the assumption that only the pump function of the diseased heart should be assisted or replaced by device while resecting the native heart is unnecessary, the concept of a "functionally total artificial heart (FTAH)" was explored. An artery pump (AP) was designed for the FTAH. The fabricated pattern of the AP, having an outer diameter of 28 mm and a length of 42 mm, was implanted in the position of ascending aorta or pulmonary trunk, joining in series to the chambers of the left ventricle or the right ventricle. The total weight of the AP pattern is 74 g, and it displaces 29 mL of volume and can achieve 5 L/min against 100 mm Hg pressure with a speed lower than 10,000 rpm. In mock circulation, two APs were connected in series with each other, and their flow rates could automatically balance each other. Simulative cardiac output and atria pressures can be maintained within a suitable range by properly adjusting the rotational speeds of each pump. Because the response of the APs to the pressure alterations at their inlets and outlets are similar to that of native heart, no intricate regulating mechanism would be necessary. This preliminary study shows that the concept of the FTAH is feasible if two APs are simultaneously implanted to replace native heart function.

Spatial genetic structure of two HIV-I-resistant polymorphisms (CCR2-64 I and SDF1-3'A) alleles in population of Shandong Province, China.

OBJECTIVE: To explore the spatial genetic structure of two HIV-I-resistant polymorphisms (CCR2-64 I and SDF1-3'A) alleles in the population of Shandong Province, China. METHODS: Using the techniques of spatial stratified sampling and spatial statistics, the spatial genetic structure of the locus (CCR2-64 I and SDF1-3'A), which was shown to be important co-receptor for HIV infection, was quantified from the populations of 36 sampled counties of Shandong Province, and a total of 3147 and 3172 samples were taken for testing CCR2-64I and SDF1-3'A respectively from individuals without known history of HIV-I infection and AIDS symptoms. RESULTS: There were significantly spatial genetic structures of the two alleles at different spatial distance classes on the scale of populations, but on the scale of individuals, no spatial structure was found in either the whole area of Shandong Province or the area of each sampled county. Although the change of frequencies of the two alleles with geographic locations in Shandong Province both showed gradual increase trends, their changing directions were inverse. The frequency of CCR2-64I allele gradually increased from the southwest to the northeast, while the frequency of SDF1-3'A allele gradually increased from the northeast to the southwest. However the RH to AIDS of combined types of their different genotypes did not represent obvious geographic diversity on the whole area of the Province. CONCLUSION: The frequency of allele usually has some spatial genetic structures or spatial autocorrelation with different spatial distance classes, but the genotypes of individuals have random distribution in the same geographic area. Evaluating spatial distribution of the genetic susceptibility of HIV (AIDS) to CCR2-64I and SDF1-3'A alleles, should focus on the frequencies of combined genotypes of CCR2 and SDF1 based on the two-locus genotypes of each individual rather than the frequencies of CCR2-64I and SDF1-3'A alleles.

The "Kriging" model of spatial genetic structure in human population genetics.

This paper presents the application of Kriging technique in the field of human population genetics for quantifying the spatial genetic heterogeneity of HLA-A locus in the area of China,and for mapping its spatial genetic structure using the measurement of synthetic genetic structure (SPC) and the principal components (PC). Both principles of the method and the basic equations are given. The Kriging model has several advantages over other interpolation and smoothing methods. Firstly, it relies on the structure of the spatial genetic semivariogram model, which can be used to quantify the spatial genetic heterogeneity of the locus (loci) before mapping its spatial genetic structure. Secondly, it is virtually unbiased in the interpolation situation,where the location to be estimated is surrounded by data on all sides and is influenced within the range of these data. Thirdly, it allows of estimative error of interpolation, which can be used to appraise the predicting effect for the spatial estimation,and the error maps can be used to decide where to introduce new sampling population genetic data. However, the "Kriging" model also has some disadvantages. Firstly,when the theoretical spatial genetic semivariogram can not be fitted by any models, the "Kriging" model can not be set up. Secondly, if the Kriging model was built by a poor spatial genetic semivariogram,the Kriging estimation standard deviation is remarkably high in the whole area, hence the Kriging model can not be suitable to estimating the distribution of spatial genetic structure. In these situations,the interpolation algorithm, whose assumption is spatial random rather than spatial autocorrelation,such as the Cavalli-Sforza method in Genography, inverse distance-weighted methods, splines, should be used to estimate or map the distribution of spatial genetic structure.